261

SKINNER,
F. A., JONES,
P. C. T.& MOLLISON,J. E. (1952). J . gen. MicroMoZ. 6,261-271.

A Comparison of a Direct- and a Plate-counting Technique

for the Quantitative Estimation of Soil Micro-organisms
BY F. A. SKINNER, P. C. T. JONES AND J. E. MOLLISON
Soil Microbiology Department, Rothamsted Ezperimental Station, Harpenden,
Hertfordshire
SUMMARY: Estimates of numbers of micro-organisms occurring in three differently
manured soils, made by a direct-counting and plating technique, were compared.
No correlation was found between the two methods and reasons for the large
discrepancy between them are discussed. Contradictory information of the effect
of external factors on soil micro-organisms can be given by the two different methods
of counting.

Estimates of the total numbers of bacteria in soil obtained by a direct-counting

technique devised by Jones & Mollison (1948) are of the order of several
thousands of millions/g. soil, while estimates by conventional plating methods
are in tens of millionslg., even on the least selective media so far used. The
same authors found a difference of the same kind between the quantity of
fungal mycelium in soil as estimated by a direct count of number of pieces/g.
as compared with colony plate counts, although in this comparison the former
count is only 2-6 times greater than the latter.
The effect of external factors such as soil moisture and temperature on the
various groups of soil microflora has been investigated by several workers and,
while certain correlations have been found, the results have been conflicting.
The estimations of numbers of organisms were mainly based on plate counts.
The present investigation was undertaken to compare direct counts with plate
counts over a period of time, to determine how these varied with soil moisture
and temperature and to show whether these two counting methods did, in
their different ways, give the same sort of information about the soilpopulation.
The soils of three differently manured plots were sampled at intervals (Fig. 1)
over the greater part of a year, and determinations made of the moisture
content of these samples and of their bacterial, fungal and actinomycetal
populations, by plating and a direct-counting technique.
'METHODS

Soil sampZing. The soil samples were taken from three of the classical plots
of Broadbalk field, which has grown wheat continuously for just over 100 years
and whose plots have had different manurial treatments over that period.
Plot 2 (farmyard manure), plot 3 (no manure) and plot 7 (complete minerals +
ammonium sulphate) were sampled at intervals between January and
December. Soil was collected by an auger to a depth of 4 in. in 12 areas on
each plot; these were bulked and passed through a 3 mm. sieve. From these
sieved samples the necessary amounts of soil were taken for the determination

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262

F . A . Skinner, P . C . T.Jones and J . E . Mollison

of moisture content, for the dilutions required for plating for the estimations
of bacteria, fungi and actinomycetes and for preparing films for direct counting.
The same samples of soil were used for estimating the numbers of amoebae
present ; these results were published separately (Singh, 1949).
Media. For plating bacteria and actinomycetes a non-selective medium,
based on the artificial soil solution used by Erikson (1947)was prepared; it
had the following composition : Ca(NO,),, 0-33g. ; CaSO,, 0-8 g. ; MgS047H20,
0.7 g.; K2S04,0.025 g.; K,HP04, 0.005 g.; NaHCO,, 0.2 g . ; distilled water to
1 1. To 1 1. of this solution were added: glucose, 0.01 g.; Difco yeast extract,
0.005 g.; FeCl,, trace; water-washed agar, 15 g. The pH was 7-2.
From the sieved soil 10 g. were taken to make the primary suspension from
which further dilutions were made in 0.75 % (w/v) NaCl. Five replicate plates
were made at a final dilution of 1/250,000using 15 ml. portions of the above
medium per plate. The plates were incubated a t 25 for 2 weeks. Although the
growth of all organisms was slow on this medium, high counts had been obtained
in preliminary tests. A total count of both bacterial and actinomycete
colonies was then made with a hand-lens. An actinomycete colony count was
made after a microscopic examination of the plates, when finely filamentous
colonies, mainly streptomyces and micromonospora, were marked. Filamentous colonies of pasty consistency (Nocardia spp.) were not counted.
For plating fungi, Czapek-Dox and Waksmans fungus-agar (Waksman,
1927, p. 19) were used; these media were adjusted to pH 4.6. Later Conns
glycerol sodium asparaginate medium was also used (Fred & Waksman, 1928) ;
it has a lower nutrient value and under test proved to be less selective and t o
give higher total counts than many other general media for fungi (Brierley,
Jewson & Brierley, 1927). Of the sieved soil 25 g. were used t o make a primary
dilution of 1/10, which was hand-shaken for 20 min. A final dilution of
1/20,000 was used for eight replicate plates having 10 ml. medium/plate.
Final counts were made after incubation for 9 days a t 25.
Direct-counting technique. Direct counts of numbers of bacteria and of the
number of pieces and total length of mycelium were estimated exactly as
described by Jones & Mollison (1948) with one modification. In the original
paper the counting of fungal mycelium with a low-power (3) objective was
advocated. In the present investigation counts were made under a Q objective
over the whole field. Although with the lower power a larger number of
observations per field is made, giving possibly greater accuracy in the final
estimation, there is some difficulty in identifying the smaller fungal fragments
and of measuring them. Higher counts are obtained using the Q objective,
as many fine filaments not resolved under a 3 objective are readily seen with
the higher power.
Soil suspensions used for the direct and plate counts were made from two
portions of the same soil sample but were prepared in different ways. For the
direct count the initial suspension was made by grinding a weighed quantity
of soil with 15 ml. distilled water, while for the plate count the initial suspension
was shaken only. This fundamental difference in technique might be considered
a source of discrepancy in the estimate of numbers of organisms, but a small

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Estimation of soil micro-organisms

experiment to compare two plate counts, one made by the usualshaken
suspension and the other by grinding the soil in the initial suspension before
dilution, showed this not to be so. Two soils, a poor soil represented by
Broadbalk plot 3, and a rich allotment sample, were used. From each sample
10 g. were used to make a primary suspension, which was shaken, and a final
dilution of 1/250,000 was used for plating. A further 2.58 g. from each soil
sample was ground up by a glass rod in 15 ml. distilled water, as in the
preliminary stages of making soil films (Jones & Mollison, 1948). One ml. was
removed to make a final dilution of 1/250,000 for plating, while the remaining
14 ml. were used for the direct-count films. The results of this experiment are
shown in Table 1, which shows that though the ground suspension gave the
higher plate count in both cases, the difference between the two plate counts
was not appreciable and the discrepancy between the plate and total direct
count was still 40-50-fold. The difference between the direct colony count and
the plate count is approximately 20-fold.
Table 1. Comparison of direct count with plate count of bacteria using two
methods for preparing soil suspensions for the plate count
Plate count
(colonies in millions/g. dry soil)
Direct count
(millions/g. dry soil)
A

Aggregates
Allotment soil
Broadbalk soil, plot 3

2220.1
1379.3

r
h

1 1

Total no.
bacteria

2 weeks
incubation

5145.6

124.4
59.8

2640.8

Ground suspension

Shaken suspension
I

5 weeks

incubation

2 weeks
incubation

5 weeks
incubation

292.8
122.0

112.3
39.5

336.9
158.2

The plates from the last experiment were kept, and since they had not dried
out were recounted after an incubation period of 5 weeks. Table 1 shows
that after 5 weeks the number of colonies had increased to two or three
times the number found after the more usual incubation period and that
the shaken suspension now gave the higher figures. The discrepancy between the direct and plate counts was here reduced approximately to one of
1O-2O-fold.
Moisture content. Samples (25 g.) were taken from the sieved soil samples
and dried overnight a t 105".
Soil temperature. The temperatures for a depth of 4 in. of soil were taken
from those of the Rothamsted meteorological records, which are made on an
area about half a mile distant from Broadbalk.
Statistical analysis. Correlation coefficients were used to assess the agreement between the numbers of organismsfg. soil estimated by the two methods
and to show whether or not these numbers were dependent on changes in soil
moisture and temperature. The correlations and regressions for the three plots
were found not to differ significantly amongst themselves and were therefore
pooled.
18

GMVI3&4

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264

F . A . Skinner, P. C. T.Jones and J . E . Mollison

RESULTS

The estimated total numbers of bacteria, fungi and actinomycetes are shown
against the sampling dates in Figs. 1 and 2. The numbers of bacteria obtained
from plate counts were, with one exception, consistently higher on plot 2 than
on plots 3 and 7,which were very similar. By the direct counting method the
total numbers of bacteria were 40-50 times higher than by the plating method.
Plot 2 again showed consistently higher numbers than plots 3 and 7 which
were again similar. It is seen from Table 2 that there is no correlation between
the two methods for bacterial counts. A partial correlation calculated after
eliminating the effect of soil moisture and temperature was still not significant.
This reflects the findings of Taylor (1936), who made a similar comparison
between plate counts and a total count obtained by the ratio method of
Thornton & Gray (1934).

Table 2. Correlation coeficients between plate a n d direct counts of bacteria,

fungi and actinomycetes
Broadbalk

Plot 2 Plot 3 Plot 7

Correlation coefficients between :

Direct bacteria and plate bacteria
Partial correlation-moisture and temperature eliminated
Direct bacteria and plate actinomycetes
Partial correlation-moisture and temperature eliminated
Plate bacteria and plate actinomycetes
Partial correlation-moisture and temperature eliminated
Direct fungi and plate fungi (Waksmans
medium)
Partial correlation-moisture and temperature eliminated
Direct fungi and plate fungi (Czapeksmedium)
Direct fungi and plate fungi (Conns medium)
Direct fungi (no. of pieces) and direct fungi
(total length)

---a47

0.082

0.392 0,342

a plots
N aggregated

-0~488 7

0-622 7

-0.286

- -0.331

0.488
0.189

D.F.

17

15
17
15

0.597

-0.320

- -0.277

17
15

0.725

0.141

0.015

0.324

23

0.089

21

-0.553

0.428

0-622

0.805 0.731

0.354 8
6
0.815 11

-0.511

0412

19

0.409

14

N =number of pairs of observations ; D.F. =degrees of freedom ;P =level of significance.

Significant correlation coefficients in black type.

The highest counts of actinomycetes by the plate method were obtained

from the plot treated with farmyard manure while the other two were very
similar, with the unmanured plot showing a tendency towards higher numbers
(Fig. 1). These figures confirm earlier work by J. Singh (1937),who found that
Broadbalk plot 2 carried a much heavier actinomycete flora than plot 3. His
figures were much lower than the present ones, probably due in part to his
having used a medium of pH 5-4, which is just above the lower limit for actinomycete development, and because in the present investigation one of us
(F.A.S.) used microscopic observation for the final identification of each
act inomycete colony.

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Estimation of soil micro-organim

265

To determine whether bacterial and actinomycete populations fluctuate in

a parallel manner, the correlation coefficient was calculated between their
respective plate counts and between the bacterial direct count and actinomycete plate count. Direct counts of actinomycetes are not possible a t present
owing to the difficulty of distinguishing their filaments from fungal hyphae
and their spores from bacteria. The aggregated correlation coefficient shows
Bacteria
Plot 2

Plot 3

Actinomycetes

24. vi.

6. viii.

Date of sampling

Fig. 1. (a) Plate counts of soil bacteria and actinomycetes in the three plots of Broadbalk
field on different dates in 1998. (Plot 2, farmyard manure; plot 3, m a n u r e d ; plot 7,
complete mineral fertilizer.) ( b ) Direct counts of soil bacteria in the same plots
as in (a).

the plate counts to be negatively and significantly correlated, whereas the

correlation between the direct bacterial count and the plate actinomycete
count is of marginal significance (P=O.1). However, when the effects of soil
moisture and temperature were eliminated, the partial correlations between
both pairs of counts showed no correlation (Table 2).
The numbers of viable fungal particles estimated in thousands/g. obtained
by plate counts of colonies on Waksmans medium are shown in Fig. 2. The
numbers found in plot 3 were consistently the lowest; plot 7 gave the highest
figure on seven of nine occasions. On Czapeks medium the numbers obtained
were of the same order; again plot 3 showed consistently the lowest figures and
plot 7 the highest on seven of eight occasions. On six of the sampling dates
(May-December) fungal counts were also made on Conns medium. The total
numbers fluctuated in a similar manner for all plots but were higher than those
18-2

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F . A . Skinner, P . C. T. Jones and J . E. Mollison

obtained on the other three media: plot 2, 280 x lo3 to 430 x lo3 colonieslg.;
plot 3, 130 x lo3 to 275 x 103/g.; plot 7, 210 x lo3 to 515 x 103/g. Plot 3 gave

266

consistently the lowest count, but plot 7 was not so consistently higher than
plot 2.
Direct counts of number of pieces of mycelium/g. of soil are shown in Fig. 2.
These estimates are from 2 to 6 times those of the plate counts. Although
plot 3 had a tendency to show the lowest figures, this method has shown less
consistent differences between the numbers on the three plots. The pieces of
mycelium observed and counted were also measured by a micrometer eyepiece and the total length of mycelium was calculated and expressed as m./g.
soil. Plot 2 showed 33-65 m., plot 3, 25-57 m., and plot 7, 25-63 m./g. soil.

$0: i.1O:ii.I
i,.iii. 2:iv. 2;.
24. i t .

IV.

24:vi.

6. biii.

29:ix.

5.ki.

8.iii.

Date of sampling

Fig. 2. Upper curves :direct counts of soil fungi expressed as number of pieces of myceliumlg.
dry soil. Lower curves: plate counts of soil fungi on Waksmans medium expressed as
number of colonies on plate/g. dry soil.

The agreement between the plate and direct methods using the figures for
number of pieces of mycelium is not good, as indicated by the correlation
coefficients (Table 2); these are positive but only barely significant for plot 2
on Waksmans medium and for plot 3 on Czapeks medium. When the
calculation is made on the aggregated observations for three plots, the correlation between the methods is just below the level of significance for all
three media. When the effects of soil moisture and temperature are eliminated
any relationship between the two counts disappears.
The two estimates of mycelial quantity, number of pieces of mycelium and
total length, parallel each other very closely, and a highly significant agreement
appears between them for every plot (Table 2). However, the counting of
pieces-an easier task than their measurement-does not necessarily give the
true picture of the development of mycelium in soil. The average length of
pieces measured in one investigation could be much higher than in another,
so that while correlation with number of pieces might be good in both, the
ratio of number of pieces :total length could be different. This did not reveal
itself in the present work, but inconsistency in the grinding of soil for making

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Estimation of soil micro-organh

soil films and differences in fragmentation of hyphal filaments could make this
a possibility.
The figures for percentage moisture content and temperature of the soil for
the dates of sampling are given in Table 3. In investigating the effect of soil
moisture and temperature on the numbers of bacteria and of fungi, it was
Table 3. Soil moisture and temperature
Moisture content (yo)
Date
20. i. 48
10.ii. 48
24. ii. 48
2. iii. 48
2. iv. 48
27. iv. 48
23. vi. 48
6.viii. 48
29. ix. 48
5. xi. 48
8.xii. 48

Plot 2
23-45
19-88
20.70
18.44
20.50
9.76
17.36
15.60
16.28
21-00
22.30

Plot 3
18.99
15-72
16.90
13-16
15.12
8-36
14-4Q
12.20
11-96
16.00
16.80

Plot 7
20.59
17.08
17-50
14.24
16.92
8.68
15.48
1340
12.60
16.80
19.00

Soil temperature
0.7'
5.5'
0.6"
3.3'
5.1'
11.3"
13.5'
17.7'
13.5'
6.8'
7.8"

Table 4. Correlation coeficients between plate and direct counts of bacteria

and soil moisture content and temperature
Broadbalk

Plot 2 Plot 3 Plot 7

orrelation coefficients between :

Direct bacteria and moisture content
Partial correlation-temperature eliminated
Plate bacteria and moisture content
Partial correlation-temperature eliminated
Direct bacteria and temperature
Partial correlation-moisture eliminated
Plate bacteria and temperature
Partial correlation-moisture eliminated

0.618 0.480
-0.309 -0.065

-0.655

0.M

0.185

-0.347 - 0 4 8

0-712 0.480
-

3 plots
N aggregated

10

0.533

0.327
-0.118
0.406

10

-0496

0.569

-0.245
0,591
0.670

D.F.

26
25
17
16
26
25
17
16

N =number of pairs of observations ; D .F.=degrees of freedom ; P =level of significance.

Significant correlation coefficients in black type.

found that direct and plate counts often indicated conflicting relations. For
instance, a positive significant correlation was found between soil temperature
and bacterial plate counts, whereas a negative significant one obtained between
soil temperature and bacterial direct counts. If the interaction of moisture be
taken into account, no correlation is then found between bacterial direct
count and temperature (Table 4). The effect of moisture on bacterial numbers
assessed by plate counting is not significant but tends to be an inverse relation, while by direct counting it is a positive and highly significant effect,
when judged by a total correlation. With the elimination of the effect of
temperature, both methods of counting show a positive and barely significant
relation with moisture. Further data on the effect of moisture and temperature
on fungi and actinomycetes are given in Table 5 . Actinomycetes show a small,
positive total correlation with moisture, significant in plot 7,and a high negative

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P
0.01
0-05-0*1

0.0.%)-1
0.01

0.01
0-01

268

P . A . Skinner, P . C . T . Jones and J . E . Mollison

correlation with temperature, significant in plot 7 and for the three plots
together. The direct fungal count also shows a significant correlation with
moisture but not for temperature, The plate counts of fungi on Czapek and
on Waksman media give total correlations with moisture and temperature
which are not very similar, although the figures for the individual plots for
each medium are fairly consistent, with the notable exception of plot 7 on
Czapek medium. A much greater number of observations would have to be
made before any explanation of these results could be offered.
Table 5 . Correlation coeflcients between fungi and actinomycetes and soil
moisture content a n d temperature
Broadbalk
A

Plot 2 Plot 3
Correlation coefficients between :
Plate actinomycetes and moisture content
Plate actinomycetes and temperature
Direct fungi and moisture
Direct fungi and temperature
Plate fungi (Waksmans medium) and moisture
Plate fungi (Czapeks medium) and moisture
Plate fungi (Waksmans medium) and temperature
Plate fungi (Czapeksmedium)and temperature

Plot 7

3 plots
N aggregated

0.278 0429 0400 7

-0.586
0.687 -0.712 7
0.590 0.544 0.418 11
-0.268 -0.139 -0404 11
0.678
0.424
0.518 9
0.273 0.695 -0.005 8
-0.127 -0.125 -0.161 9
0.356

0.561

0.587

8 .

D.F.

P
0.054.1
0.01

0.550
0.142

17
17
29
29
23
19

0.453

19

0.02-0-05

0.448

-0,633
0.517

-0.266

0.01

0.01

N =number of pairs of observations; D.F. =degrees of freedom ;P =level of significance.

Significant correlation coefficients in black type.

DISCUSSION

In the present investigation direct counts of numbers of bacteria and fungi

were always much higher than those of corresponding plate counts, and the two
methods gave different information about their fluctuations. Several factors
may contribute to the large discrepancy in numbers given by the two methods :
(1) clumps of bacteria which remain aggregated in spite of the shaking that
precedes plating; (2) staining of non-viable particles in direct count preparations ; (3) failure to distinguish between bacteria and actinomycete spores,
and between fungal and actinomycete filaments in stained films; (4) competition on plates; ( 5 ) selectivity of plating media and non-cultivation of obligate
anaerobes.
The frequency with which plate colonies develop from aggregates of bacteria
rather than from individual cells is not easy to assess but there is strong
evidence that this must occur. It is shown in Table 6 that the number of
observations of cells in colonies of two or more was about the same as that
of single cells. These small aggregates had resisted grinding in the preparation
of the soil films, and it is unlikely that similar ones would have disintegrated
into their component cells during the shaking that preceded plating. If, as is
probable, aggregates such as those observed in soil films gave rise to only
single colonies on plates, this alone would have made the direct count approximately double the plate count. Further, a minute soil particle bearing several

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Estimation of soil micro-organisms

269

individuals or even aggregates of different species might give only one colony
on a plate. The occurrence of impure colonies on plates suggests strongly that
this may frequently happen.
Table 6. Analysis of bacterial aggregates occurring on soil films
No. aggregates per slide
r

Broadbalk (plot 3)
Slide no.:
No. of cells
in aggregate

62 49
1
35 30
2
4
6
3
9 1 2
4
4
2
5
2
3
6
1
7
2
8
9
- 10
- 11
12
14
16
17
20
21
53
Total no. of 117 104
aggregates
Total no. of 219 219
cells
Aggregates
Ratio :
0.53 0.47
Total cells

61
31
3
2
3
1
2

63
16
6
3
2
2
2
-

104

96

Allotment
1
1
2
3
4

85
31
7
9
2
6
1
2
1

82
32
6
15

7
4

82
34
8
11
9
5
1
1

89
48
5
15
3
5
1
4

187 181 339 348 360 397

0.56 0.53 0.43 0.43 043 0.44

The staining of dead cells in direct count preparations may be expected

to introduce a serious error. That many hyphal pieces observed under the
microscope seem empty of protoplasmic contents and are purplish in colour
instead of the intense blue shown by known actively growing mycelium was
pointed out by Jones & Mollison (1948). To a lesser degree bacterial cells of
varying intensity of blue can be seen, but to attempt to separate viable from
non-viable particles by depth of colour would be too arbitrary a procedure.
It is interesting to note that Taylor (1936), in experiments with soil cultures
of bacteria (all of which were able to grow on a soil extract agar), found a plate
count on this same medium to be 36-61 % of a direct count estimated by the
ratio technique of Thornton & Gray (1934), which gives counts of a magnitude
similar to that given by the method used in the present work. As shown in
Table 6 a difference of 36-61 % can be accounted for if clumps of several cells
develop into single colonies on plates, and thus cannot necessarily be attributed
to the staining of non-viable cells. On the other hand, the number of non-

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270

F . A . Skinner, P . C. T.Jones and J . E . Mollison

viable but stainable cells may accumulate with time and may therefore be
greater in a field soil than in an artificial soil population incubated for the
relatively short periods used by Taylor. At present, the separation of viable
from non-viable organisms in direct counting remains impracticable.
The difficulty of distinguishing bacteria from actinomycete spores is well
known. These spores abound in soils, and there is strong evidence that in some
soils actinomycetes may be present largely in this form (Skinner, 1951).
Allowing that some actinomycetes may not develop on plates, it seems
possible that almost the whole actinomycete count of 10-15 millionlg. soil is
included in the direct bacterial count. However, unless there are vast numbers
of dead actinomycete spores present, this source of error cannot be a large one
as the actinomycete plate count is small compared with the direct bacterial
count.
Competition between colonies on plates and the antagonistic suppression
of one species by another are factors which may cause a lower plate count,
but no evidence can be presented to show that they were operating in the
present investigation.
It has been shown that the growth of single colonies from bacterial aggregates would account for a plate count only half the corresponding direct
count. However, this alone is inadequate to explain the enormous discrepancy
between numbers given by the two methods. The most probable further
causes of this discrepancy would seem to be the counting of non-viable particles
in soil films (tending to increase the direct count) and selectivity of the plating
medium, a well-known limitation of all plating techniques tending to reduce
the plate count.
The ratio fungal plate count : direct count is only 1 : 2-6; selectivity of
the plating medium and the direct counting of non-viable fragments could
explain this small differencesatisfactorily. In view of Skinners (1951) results,
it seems unlikely that actinomycete fragments contribute appreciably to the
fungal direct count. It is important to remember that different soil conditions
may encourage sporulation of fungi rather than mycelial growth. For this reason
a plate count may give a false picture of mycelial development while a direct
count may give a truer analysis.
Many studies on the effect of external factors on the fluctuating numbers of
micro-organisms have been made, and although the results have often been
contradictory, most workers consider that variations in soil moisture content
must have some effect on the micro-population, although Taylor (1936) showed
that fluctuations of bacterial numbers took place while soil moisture content
and temperature were kept constant. It is evident that neither direct nor
plate counting records accurately changes in the total numbers of soil microorganisms. In direct counts such fluctuations may be masked by an accumulation of non-viable but stainable cells. The selectivity of plating media and
the fact that here plate counts of bacteria were positively and direct counts
negatively correlated with temperature suggest that the plate count is an
estimate of one element of the soil population, which reacts to external factors
differently from the total population. With these considerations in mind it is

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Estimation of soil micro-organisms

271

easier to understand the absence of correlations between plate and direct

counts.
Plate counting is a well-established method, perhaps of greatest value for
qualitative investigation. We consider that the new direct-counting technique,
though it undoubtedly gives an overestimate of the viable population, has
proved reliable for quantitative investigations. It is rapid, and as both
bacteria and fungi can be examined in the same soil film, it saves labour and
equipment necessary for plating experiments.
Two of us (F.A.S. and J.E.M.) held grants from the Agricultural Research Council
when this work was undertaken. We wish to record our thanks to Dr H. G. Thornton,
F.R.S., for his helpful advice, to J. H. A. Dunwoody for assistance with the statistical
work, and to Miss Mabel Dunkley for preparation of the typescript.
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(Received 30 August 1951)

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